杂交竹抗梢枯病诱导因子筛选及其持续期研究
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摘要
本文利用病原毒素同源异质的特点诱导杂交竹抗病潜力,筛选出抗梢枯病的最佳诱导因子并排除其对病原菌的直接作用,经琼脂玻片萌发法测定3种诱导因子(温度灭活毒素、蛋白酶降解毒素和细胞壁成分)对杂交竹梢枯病病原菌暗孢节菱孢菌孢子萌发的抑制作用,结果显示经过60℃灭活毒素浓度为40μg/mL处理的孢子萌发效果最为理想。通过最佳诱导因子对杂交竹不同品种诱导持续期进行测定,采用针刺法先接种诱导因子后挑战接种病原菌,1～40 d内观察抗感品种对诱导因子响应的差异,症状上杂交竹8#的感病程度重于3#和6#,40 d时叶片和枝干变黄干枯,病情指数结果表明,3个杂交竹品种接种诱导因子后感病程度降低,诱抗效果显示杂交竹6#的诱抗指数高于3#和8#。以上结果证明诱导因子使不同杂交竹品种均产生了一定的抗性且抗性越强的品种诱导抗性越好。
By the homologous heterogeneity of pathogenic toxins, the resistance potential of Bambusa pervariabilis×Dendrocalamopisis grandis was induced; the optimum inducing factor was screened and the direct effect was excluded. The inhibitory effects of three factors(inactivated toxin by temperature, degraded toxin by protease and cell wall component) on spore germination of Arthrinium phaeospermum, the pathogen of B.pervariabilis×D.grandis blight, were determined by agar slide germination method. The results showed that the spore germination effect of the toxin treated at 60℃ at the concentration of 40 μg/mL was the best. The induction durations of different varieties of B.pervariabilis×D.grandis were determined through the optimum inducing factor; the needling method was used to inoculate the inducing factor first and then the pathogen within 40 days to observe the response difference between the resistant and susceptible varieties to the inducing factor. The results showed that B.pervariabilis×D.grandis #8 was more susceptible than #3 and #6, with the yellowish and withered leaves and branches at 40 d. The disease indices of the three varieties were decreased after inoculation of the inducing factor, and the induced resistance effect of B.pervariabilis×D.grandis #6 was higher than those of #3 and #8. These results indicated that the inducing factor could induce certain resistance in different varieties of B.pervariabilis×D.grandis, and the stronger the original resistance, the better the induced resistance.
By the homologous heterogeneity of pathogenic toxins, the resistance potential of Bambusa pervariabilis×Dendrocalamopisis grandis was induced; the optimum inducing factor was screened and the direct effect was excluded. The inhibitory effects of three factors(inactivated toxin by temperature, degraded toxin by protease and cell wall component) on spore germination of Arthrinium phaeospermum, the pathogen of B.pervariabilis×D.grandis blight, were determined by agar slide germination method. The results showed that the spore germination effect of the toxin treated at 60℃ at the concentration of 40 μg/mL was the best. The induction durations of different varieties of B.pervariabilis×D.grandis were determined through the optimum inducing factor; the needling method was used to inoculate the inducing factor first and then the pathogen within 40 days to observe the response difference between the resistant and susceptible varieties to the inducing factor. The results showed that B.pervariabilis×D.grandis #8 was more susceptible than #3 and #6, with the yellowish and withered leaves and branches at 40 d. The disease indices of the three varieties were decreased after inoculation of the inducing factor, and the induced resistance effect of B.pervariabilis×D.grandis #6 was higher than those of #3 and #8. These results indicated that the inducing factor could induce certain resistance in different varieties of B.pervariabilis×D.grandis, and the stronger the original resistance, the better the induced resistance.
引文
[1] 赵继红, 孙淑君, 李建中. 植物诱导抗病性与诱抗剂研究进展[J]. 植物保护, 2003, 29(4): 7-10.
[2] YE X S, PAN S Q, KUC J. Activity isozyme pattern, and cellular localization of peroxidase as related to systemic resistance of tobacco to blue mold (Peronospora tabacina) and to Tobacco mosaic virus [J].Phytopathology,1990,80(12):1295-1299.
[3] 赵继红,李建中.灰霉菌菌丝体提取物诱导番茄抗病性的初步研究[J].河南农业科学,2003(10):55-56.
[4] 胡景江,刘志龙,文建雷.溃疡病菌低聚糖激发子诱导杨树细胞抗病机制的初步研究[J].西北农林科技大学学报(自然科学版),2003,31(4):145-148.
[5] 李云锋,王振中,贾显禄.稻瘟菌激发子CSBI诱导水稻防御性相关酶的活性变化[J].作物学报,2004,30(6):613-617.
[6] 李洪连,王守正,王金生,等.诱抗菌激发子诱导黄瓜产生对炭疽病的抗性[J].植物生理学报,1993,19(4):320-324.
[7] 郭红莲,陈捷,高增贵,等.不同诱抗剂诱导玉米对灰斑病的抗性及其与PAL的关系[J].沈阳农业大学学报,2000,31(5):465-467.
[8] 宾金华,潘瑞炽.茉莉酸甲酯对烟草幼苗抗炭疽病的影响[J].热带亚热带植物学报,1997,5(4):21-26.
[9] 宾金华,潘瑞炽.茉莉酸甲酯诱导烟草幼苗抗病与过氧化物酶活性和木质素含量的关系[J].应用与环境生物学报,1999,5(2):160-164.
[10] 宾金华,姜胜,黄胜琴,等.茉莉酸甲酯诱导烟草幼苗抗炭疽病与PAL活性及细胞壁物质的关系[J].植物生理学报,2000,26(1):1-6.
[11] KUBO M, SATO T. Utilization of high temperature stress as plant resistance activators for control of summer greenhouse cucumber diseases [J].Acta Horticulturae,2002,588:171-174.
[12] 李宝聚,彭霞薇,王福建,等.高温诱导黄瓜抗黑星病研究[J].园艺学报,2001,28(2):177-179.
[13] DEAN R A, KUC J. Rapid lignification in response to wounding and infection as a mechanism for induced systemic protection in cucumber [J]. Physiological and Molecular Plant Pathology, 1987, 31:69-81.
[14] 朱天辉,黄宗超,高强章,等.撑X绿杂交竹枯梢病病原及发生规律研究[J].中国森林病虫,2009,28(2):10-12.
[15] 李姝江.杂交竹梢枯病菌蛋白毒素及其精确作用机制研究[D].成都:四川农业大学,2013.
[16] LI S J, ZHU T H. Purification of the toxin protein Pc from Arthrinium phaeospermum and its effect on the defence enzymes of Bambusa pervariabilis×Dendrocalamopsis grandis varieties [J]. Forest Pathology, 2014,44(2): 96-106.
[17] LI S J, ZHU T H, ZHU H M Y, et al. Purification of protein AP-toxin from Arthrinium phaeospermum causing blight in Bambusa pervariabilis×Dendrocalamopsis grandis and its metabolic effects on four bamboo varieties [J]. Phytopathology, 2013, 103(2): 135-145.
[18] 黄昌华.用琼脂玻片法测定孢子萌发率[J],植物保护,1993(3):36-37.
[19] 卢丹,徐晴,江凌,等.生物酶降解真菌毒素的研究进展[J].生物加工过程,2018,16(2):49-56.
[20] LI S J, ZHU T H. Biochemical response and induced resistance against anthracnose (Colletotrichum camelliae) of camellia (Camellia pitardii) by chitosan oligosaccharide application[J]. Forest Pathology, 2013, 43:67-76.
[21] 方中达.植病研究方法[M].北京:中国农业出版社,1998.
[22] 黄艳娜.外源性水杨酸诱导山茶对灰斑病抗性的研究[D].成都:四川农业大学,2006.
[23] 葛银林,李德葆.植物抗病性的诱导、机制、分子生物学研究进展[J].中国生物防治,1995,11(3):134-141.
[24] 李洪连,王守正,袁红霞,等.植物诱导抗病性研究的现状与展望[J].河南农业大学学报,1994,28(3):219-229.
[25] 李洪连,王守正,王金生,等.黄瓜对炭疽病诱导抗性的初步研究Ⅱ.诱抗抗病机制的研究[J].植物病理学报,1993,23(4):327-332.
[26] 李开本,陈福如,翁启勇,等.木霉菌对黄瓜生长及过氧化物酶影响初报[C]//中国植物保护研究进展.北京:中国科学技术出版社,1996:288-292.
[27] 李开本,翁启勇,陈庆河,等.健株作物根际微生物优势种对黄瓜生长影响初报[C]//中国植物保护研究进展.北京:中国科学技术出版社,1996:285-287.
[28] MAYAMA S, TANI T, UENO T, et al. The purification of victorin and its phytoalexin elicitor activity in oat leaves [J]. Physiological and Molecular Plant Pathology, 1986, 29: 1-8.
[29] EHRENSHAFT M,UPCHURCH R G. Host protein(s) induces accumulation of the toxin cercosporin and mRNA in a phytopathologenic strain of Cercospora kikuchii [J]. Physiological and Molecular Plant Pathology, 1993, 43(2): 95-107.
[30] MOUSSATOS V V, YANG S F, WARD B,et al. AAL-toxin induced physiological changes in Lycopersicon esculentum Mill: roles for ethylene and pyrimidine intermediates in necrosis [J]. Physiological and Molecular Plant Pathology, 1994, 44(6): 455-468.
[31] 章元寿. 植物病原真菌毒素的研究现状[J]. 真菌学报, 1991, 10(3): 169-181.
[32] 董金皋. 玉米大斑病菌HT-毒素Ⅰ的提纯、结构鉴定及致病活性研究[D]. 北京:中国农业大学, 1998.
[33] OKUNO T, NAKAYAMA M, OKAJIMA N, et al. Systemic resistance to downy mildew and appearance of acid soluble proteins in cucumber leaves treated with biotic and abiotic inducers[J]. Annals of the Phytopathological Society of Japan,2009, 57(2):203-211.
[34] 冉隆贤,谷文众,吴光金.水杨酸诱导桉树抗青枯病的作用及相关酶活性变化[J].林业科学研究,2004,17(1):12-18.
[35] 蔡新忠,郑重,宋凤鸣.水杨酸对水稻幼苗抗瘟性的诱导作用[J].植物病理学报,1996,26(1):7-12.
[36] 柳建良,黄小丹,傅炽栋,等.水杨酸对芒果炭疽病的诱导抗性作用[J].热带亚热带植物学报,1998,6(3):245-248.
[37] 李红玉,郭金魁,周功克.水杨酸诱导黄瓜抗黑星病抗性的部位差异和时效性[J].应用与环境生物学报,1999,5(6):640-642.
[38] 刘亚光.大豆灰斑病菌毒素组分、致病性及其诱导抗性的研究[D].哈尔滨:东北农业大学,2002.
[39] 王敬文,薛应龙.植物苯丙氨酸解氨酶的研究——Ⅱ苯丙氨酸解氨酶在抗马铃薯晚疫病中的作用[J].植物生理学报,1982,8(1):35-42.
[2] YE X S, PAN S Q, KUC J. Activity isozyme pattern, and cellular localization of peroxidase as related to systemic resistance of tobacco to blue mold (Peronospora tabacina) and to Tobacco mosaic virus [J].Phytopathology,1990,80(12):1295-1299.
[3] 赵继红,李建中.灰霉菌菌丝体提取物诱导番茄抗病性的初步研究[J].河南农业科学,2003(10):55-56.
[4] 胡景江,刘志龙,文建雷.溃疡病菌低聚糖激发子诱导杨树细胞抗病机制的初步研究[J].西北农林科技大学学报(自然科学版),2003,31(4):145-148.
[5] 李云锋,王振中,贾显禄.稻瘟菌激发子CSBI诱导水稻防御性相关酶的活性变化[J].作物学报,2004,30(6):613-617.
[6] 李洪连,王守正,王金生,等.诱抗菌激发子诱导黄瓜产生对炭疽病的抗性[J].植物生理学报,1993,19(4):320-324.
[7] 郭红莲,陈捷,高增贵,等.不同诱抗剂诱导玉米对灰斑病的抗性及其与PAL的关系[J].沈阳农业大学学报,2000,31(5):465-467.
[8] 宾金华,潘瑞炽.茉莉酸甲酯对烟草幼苗抗炭疽病的影响[J].热带亚热带植物学报,1997,5(4):21-26.
[9] 宾金华,潘瑞炽.茉莉酸甲酯诱导烟草幼苗抗病与过氧化物酶活性和木质素含量的关系[J].应用与环境生物学报,1999,5(2):160-164.
[10] 宾金华,姜胜,黄胜琴,等.茉莉酸甲酯诱导烟草幼苗抗炭疽病与PAL活性及细胞壁物质的关系[J].植物生理学报,2000,26(1):1-6.
[11] KUBO M, SATO T. Utilization of high temperature stress as plant resistance activators for control of summer greenhouse cucumber diseases [J].Acta Horticulturae,2002,588:171-174.
[12] 李宝聚,彭霞薇,王福建,等.高温诱导黄瓜抗黑星病研究[J].园艺学报,2001,28(2):177-179.
[13] DEAN R A, KUC J. Rapid lignification in response to wounding and infection as a mechanism for induced systemic protection in cucumber [J]. Physiological and Molecular Plant Pathology, 1987, 31:69-81.
[14] 朱天辉,黄宗超,高强章,等.撑X绿杂交竹枯梢病病原及发生规律研究[J].中国森林病虫,2009,28(2):10-12.
[15] 李姝江.杂交竹梢枯病菌蛋白毒素及其精确作用机制研究[D].成都:四川农业大学,2013.
[16] LI S J, ZHU T H. Purification of the toxin protein Pc from Arthrinium phaeospermum and its effect on the defence enzymes of Bambusa pervariabilis×Dendrocalamopsis grandis varieties [J]. Forest Pathology, 2014,44(2): 96-106.
[17] LI S J, ZHU T H, ZHU H M Y, et al. Purification of protein AP-toxin from Arthrinium phaeospermum causing blight in Bambusa pervariabilis×Dendrocalamopsis grandis and its metabolic effects on four bamboo varieties [J]. Phytopathology, 2013, 103(2): 135-145.
[18] 黄昌华.用琼脂玻片法测定孢子萌发率[J],植物保护,1993(3):36-37.
[19] 卢丹,徐晴,江凌,等.生物酶降解真菌毒素的研究进展[J].生物加工过程,2018,16(2):49-56.
[20] LI S J, ZHU T H. Biochemical response and induced resistance against anthracnose (Colletotrichum camelliae) of camellia (Camellia pitardii) by chitosan oligosaccharide application[J]. Forest Pathology, 2013, 43:67-76.
[21] 方中达.植病研究方法[M].北京:中国农业出版社,1998.
[22] 黄艳娜.外源性水杨酸诱导山茶对灰斑病抗性的研究[D].成都:四川农业大学,2006.
[23] 葛银林,李德葆.植物抗病性的诱导、机制、分子生物学研究进展[J].中国生物防治,1995,11(3):134-141.
[24] 李洪连,王守正,袁红霞,等.植物诱导抗病性研究的现状与展望[J].河南农业大学学报,1994,28(3):219-229.
[25] 李洪连,王守正,王金生,等.黄瓜对炭疽病诱导抗性的初步研究Ⅱ.诱抗抗病机制的研究[J].植物病理学报,1993,23(4):327-332.
[26] 李开本,陈福如,翁启勇,等.木霉菌对黄瓜生长及过氧化物酶影响初报[C]//中国植物保护研究进展.北京:中国科学技术出版社,1996:288-292.
[27] 李开本,翁启勇,陈庆河,等.健株作物根际微生物优势种对黄瓜生长影响初报[C]//中国植物保护研究进展.北京:中国科学技术出版社,1996:285-287.
[28] MAYAMA S, TANI T, UENO T, et al. The purification of victorin and its phytoalexin elicitor activity in oat leaves [J]. Physiological and Molecular Plant Pathology, 1986, 29: 1-8.
[29] EHRENSHAFT M,UPCHURCH R G. Host protein(s) induces accumulation of the toxin cercosporin and mRNA in a phytopathologenic strain of Cercospora kikuchii [J]. Physiological and Molecular Plant Pathology, 1993, 43(2): 95-107.
[30] MOUSSATOS V V, YANG S F, WARD B,et al. AAL-toxin induced physiological changes in Lycopersicon esculentum Mill: roles for ethylene and pyrimidine intermediates in necrosis [J]. Physiological and Molecular Plant Pathology, 1994, 44(6): 455-468.
[31] 章元寿. 植物病原真菌毒素的研究现状[J]. 真菌学报, 1991, 10(3): 169-181.
[32] 董金皋. 玉米大斑病菌HT-毒素Ⅰ的提纯、结构鉴定及致病活性研究[D]. 北京:中国农业大学, 1998.
[33] OKUNO T, NAKAYAMA M, OKAJIMA N, et al. Systemic resistance to downy mildew and appearance of acid soluble proteins in cucumber leaves treated with biotic and abiotic inducers[J]. Annals of the Phytopathological Society of Japan,2009, 57(2):203-211.
[34] 冉隆贤,谷文众,吴光金.水杨酸诱导桉树抗青枯病的作用及相关酶活性变化[J].林业科学研究,2004,17(1):12-18.
[35] 蔡新忠,郑重,宋凤鸣.水杨酸对水稻幼苗抗瘟性的诱导作用[J].植物病理学报,1996,26(1):7-12.
[36] 柳建良,黄小丹,傅炽栋,等.水杨酸对芒果炭疽病的诱导抗性作用[J].热带亚热带植物学报,1998,6(3):245-248.
[37] 李红玉,郭金魁,周功克.水杨酸诱导黄瓜抗黑星病抗性的部位差异和时效性[J].应用与环境生物学报,1999,5(6):640-642.
[38] 刘亚光.大豆灰斑病菌毒素组分、致病性及其诱导抗性的研究[D].哈尔滨:东北农业大学,2002.
[39] 王敬文,薛应龙.植物苯丙氨酸解氨酶的研究——Ⅱ苯丙氨酸解氨酶在抗马铃薯晚疫病中的作用[J].植物生理学报,1982,8(1):35-42.